anatomical and phytochemical study of plant parts of butea...

Int. J. of Life Sciences, 2016, Vol. 4 (2): 227-234 ISSN: 2320-7817| eISSN: 2320-964X

© 2016 |IJLSCI www.ijlsci.in 227

Anatomical and Phytochemical study of plant parts of Butea

monosperma (Lamk.) Taub.

Gupta Pooja1*, Chandra Naresh2 and Bhot Meeta3

1Department of Botany, Birla College, 2Principal, Birla College, 3Associate Professor, Department of Botany 1,2,3Birla College of Arts, Science and Commerce, Kalyan (W), Maharashtra – 421304.

*Corresponding author email:poojaog84@gmail

Manuscript details: ABSTRACT

Received: 31.03.2016 Accepted: 10.06.2016 Published : 23.07.2016 Editor: Dr. Arvind Chavhan Cite this article as: Gupta Pooja, Chandra Naresh and

Bhot Meeta (2016) Anatomical and

Phytochemical study of plant parts

of Butea monosperma (Lamk.) Taub,

International J. of Life Sciences, 4(2):

227-234.

Copyright: © 2016 | Author(s), This is an open access article under the terms of the Creative Commons Attribution-Non-Commercial - No Derivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

The plant kingdom is a treasure house of potential drugs and in the

recent years there has been an increasing awareness about the

importance of medicinal plants. Drugs from the plants are easily

available, less expensive, safe and rarely have side effects. The plants

which have been selected for medicinal use over thousands of years

constitute for examining the current search for therapeutically effective

new drugs such as anticancer drugs, antimicrobial drugs, anti-

hepatotoxic compounds. Butea monosperma (Lamk.) Taub. is one such

medicinal plant commonly known as palas (Flame of the forest)

belonging to the family Fabaceae distributed throughout India.It is

imperative that any crude drug for pharmacological use, needs to be

subjected to scrutiny for botanical identity. The role of anatomical and

phytochemical analysis is sought at this juncture to provide a set of

diagnostic features of the plant which will help to a considerable extent

to ascertain the botanical identification of the drugs. The present study

has been carried out to determine the anatomical, phytochemical

screening and HPTLC finger printing analysis of different parts of Butea

monosperma (Lamk.) Taub. was carried out for crude drug identification

and for standardization of the quality.

Keywords: Butea monosperma, anatomy, herbal medicine,

morphological study, microscopical study, phytochemical study

INTRODUCTION

Butea monosperma (Lamk.) Taub. is a medicinal plant commonly known

as palas (Flame of the forest) belonging to the family Fabaceae

distributed throughout India, except arid parts (Sharmaetal., 2000).

Different species of Butea found in India are Butea parviflora,Butea

purpurea, Butea monosperma and Butea superb (Almeida, 1998). It is said

that this treeisa form of Agnidev, Godof Fire. Butea monosperma (Lamk.)

Taub. has various healing effects which are seen in treatment of many

diseases. The bark is an appetiser, lessens inflammation, used in liver

RESEARCH ARTICLE

Gupta et al., 2016

228 Int. J. of Life Sciences, Vol. 4(2) June, 2016

disorders, fractures, topically in piles and purifies the

blood. The leaf is an appetiser, astringent,

anthelmintic, aphrodisiac, cures boils and piles. The

flowers are astringent, diuretic and aphrodisiac and

they are used to disperse swellings. The fruit and the

seeds are bitter and oily and useful in piles, eye

diseases and inflammation (Kirtikar and Basu, 1984).

In the present work, the morphological, microscopical

and phytochemical studies of different parts of Butea

monosperma (Lamk.) Taub. have been carried out

which will be useful for proper identification and

authentication of crude drug.

MATERIALS AND METHODS

Butea monosperma (Lamk.) Taub. plant parts (young

stem, old stem, bark, leaf, and petiole) were collected.

Plant was identified and authenticated from Blatter

Herbarium St. Xavier’s College, Mumbai.The collected

plant parts (leaf, bark and flowers) were dried under

shade and powdered with mechanical grinder and

stored in air tight containers and used for the study.

Anatomical study

Anatomical features were studied by taking transverse

sections of young and old stem (T.L.S, R.L.S of stem),

leaf and petiole of Butea monosperma (Lamk.) Taub.

sections were stained by safranine and mounted in

glycerine and observed under microscope at 10X (low

magnification power) and 45X (high magnification

power). Different wood elements were studied using

maceration technique and by powder analysis.

Phytochemical analysis

The preliminary phytochemical analysis for plant parts

(leaf, bark and flowers) of Butea monosperma (Lamk.)

Taub. was carried out using standard methods

(Sofowra, 1993; Trease and Evans, 1989 and

Harborne, 1973). The quantitative analysis of

secondary metabolites such as flavonoids, saponins

and phenols of different parts were also carried using

standard methods. HPTLC fingerprint confirmed the

presence of flavonoids and saponins in leaf, bark and

flowers of Butea monosperma (Lamk.) Taub. The

separation of flavonoids was achieved using ethyl

acetate-formic acid-glacial acetic acid-water

(100:11:11:26) and for saponins using mobile phase

chloroform-acetic acid-methanol-water (64:32:12:8)

(Wagner and Bladt., 1986).

RESULTS AND DISCUSSION

Anatomical study

Morphological study

Butea monosperma(Lamk.) Taub.is a medium sized

erect tree (12-15m) with crooked trunk and irregular

branches (Figure 1.1). Bark is rough and ash coloured.

Leaves are trifoliate (10-20cm long and broad), with

Figure1.2: Leaves

Figure 1.1: Tree Figure1.3: Flowers

Anatomical and Phytochemical study of plant parts of Buteamonosperma (Lamk.) Taub

www.ijlsci.in Int. J. of Life Sciences, Vol. 4(2) June, 2016 229

long petiole and stipulate (Figure 1.2). Leaflets are

coriaceous. All leaflets are obtuse, glabrous when old,

finely silky and with reticulate venation. Flowers are

large and produced in rigid racemes (15cm long)

(Figure 1.3).

Three flowers together form the tumid nodes of the

dark olive-green velvety rachis and pedicels are about

twice as long as the calyx and densely brown-velvety.

Bracts and bracteoles are small and deciduous. Calyx is

long, dark olive-green, densely velvety on outside,

clothed with silky hairs within teeth, the upper two are

connate and the lower three are equal and deltoid.

Corolla (3.5-5cm long) clothed outside with silky,

silver hairs and is orange or salmon coloured with

broad standard and semi-circular keel, beaked and

veined. Pods are stalked, thickened at the sutures and

reticulately veined.

Microscopical study

T.S of old stem of Butea monosperma (Lamk.) Taub.

Old stem of Butea monosperma (Lamk.) Taub.is wavy

in outline. Old stem shows both extrastelar and

intrastelar secondary growth. Extrastelar secondary

growth gives rise to periderm made up of phellem,

phellogen and phelloderm thus forming a bark.

Intrastelar secondary growth shows development of

wood (secondary xylem) which is of diffused porous

type. In wood vessels are scattered and at some places

they are arranged in long radial multiples. Number of

vessels ranges from 1-4 in each radial cluster. Wood

parenchyma is of paratracheal type where the

parenchyma is in direct association with vessels.

Secondary phloem shows presence of yellow gummy

mass scattered here and there. Medullary rays are

formed for radial conduction. Bundle caps of primary

Figure 1.4: T.S. of old stem Figure 1.5: T.S. of old stem

A - Entire view, B- Secondary growth view Keywords: Ygm- Yellow gummy mass; Xv - Xylem vessels;Bc- Bundle cap;

Mr - Medullary rays; T - Tracheid; P –Periderm

Figure 1.6: T.S. of young stem Figure 1.7: T.L.S. of stem

Keywords: Ygm- Yellow gummy mass; Xv - Xylem vessels;Bc- Bundle cap;

Mr - Medullary rays; T - Tracheid; P -Perider

Gupta et al., 2016


Figure 1.8: R.L.S. of stem

Figure 1.9: Powder characteristics of old stem of Butea monosperma(Lamk.)

Keywords:A – Vessel; B – Ca-oxalatecrystalsand yellow gummy mass;C – Tracheid;

D – Tracheid with annular thickening; E – Pitted vessels; F – Reticulate thickening

vascular bundle are pushed towards periphery.

Parenchymatous pith is at the center with deposition

of yellow gummy mass (Figure1.4 and 1.5).

T.S of young stem of Butea monosperma (Lamk.)

Taub.

Young stem of Butea monosperma (Lamk.) Taub.is

slightly wavy in outline. Cortex is made up of two types

of tissues, sclerenchyma forming outer cortex and

parenchyma in inner cortex. Few innermost cells of

inner cortex show chlorenchymatous tissue just above

the bundle caps. Vascular bundles are arranged in ring

with wavy outline. Each vascular bundle is conjoint,

collateral and open with endarch xylem. Phloem

region is broad with deposition of gummy mass. Stele

siphonostele, as large parenchymatous pith is at the

center which also shows deposition of gummy mass

(Figure 1.6).

T.L.S of stem

T.L.S of stem of Butea monosperma(Lamk.) Taub.

shows long uniseriate and multiseriate rays made up

of parenchymatous cells (Figure 1.7).

R.L.S of stem

R.L.S of stem of Butea monosperma (Lamk.) Taub.

shows xylem with tracheids and vessels. Radial wall of

tracheids shows annular thickening. Vessels are with

pitted and reticulate thickening. Other elements like

xylem parenchyma and fibers are also found in traces

(Figure 1.8)



Powder of old stem after maceration shows vessels

with pits on their lateral walls, tracheids with annular

thickening, vessels with reticulate thickening, yellow

gummy mass and Ca-oxalate crystals (Figure 1.9).

T.S of leaf

Leaf of Butea monosperma(Lamk.) Taub. shows upper

and lower epidermis with unicellular pointed hairs

from their surface. Leaf lamina shows dorsiventral

structure with palisade and spongy tissue. In midrib

region, heterogeneous cortex was observed with

sclerenchymatous and chlorenchymatous tissue. In the

midrib vascular bundles are in ring with bundle caps

on their outer side. Vascular bundles are conjoint and

collateral. Center of midrib shows parenchyma with

deposits of yellow gummy mass (Figure 1.10).

Powder of leaf shows presence of unicellular pointed

hairs, vessels with wall thickening and yellow gummy

mass (Figure 1.12 A-D).

Figure 1.10: T.S. of leaf Figure 1.11: T.S of petiole

Figure 1.12: Powder characteristics of leaf of Butea monosperma(Lamk.) Taub.

Keywords:A – Vessel; B – Scalariformvessel; C - Yellow gummy mass; D – Unicellular hair

Gupta et al., 2016


Table 1.1: Preliminary phytochemical analysis of Butea monosperma (Lamk.) Taub. leaf, bark and flowers.

Phyto- Constituents

Method/Test Plant parts

L B F

Triterpenes Salkowski’s test + - +

Phenol Ferric Chloride test + + +

Tannins Ferric Chloride test + + +

Proteins Xanthoproteic test + - +

Flavonoids Ammonium test + + +

Alkaloids Mayer’s reagent and Wagner’s reagent test - + +

Saponins Froth Emulsion test + + +

Keys: + = Present; - = Absent; L = Leaf; B = Bark; F = Flower

Table 1.2: Quantitative analysis of Butea monosperma(Lamk.) Taub. leaf, bark and flower.

Values are Mean±S.D. of three determinations

Keys: + = Present; - = Absent; L = Leaf; B = Bark; F = Flowers

T.S. of petiole

T.S of petiole of Butea monosperma (Lamk.) Taub.

showed circular outline. Internally it is differentiated

into epidermis, cortex and stele. Hairs are unicellular

and pointed arising from surface of epidermis. Outer

cortex is 1-3 layered and sclerenchymatous. While

inner cortex is broad and parenchymatous. Stele is

horse shoe or wedge shaped in outline. Vascular

bundles are with sclerenchymatous cap on the outer

side and are conjoint and collateral. Xylem vessels are

arranged in radial clusters and are of endarch type.

Stele is siphonostele (Figure 1.11).

Phytochemical analysis

Preliminary Qualitative analysis

Preliminary qualitative estimation of phytochemicals

in powdered plant parts (leaf, bark and flowers) is

summarized in (Table 1.1). Leaf, bark and flowers

powder of Butea monosperma (Lamk.) Taub. showed

presence of flavonoids, saponins, phenols and tannins.

The powdered leaf and flowers of Butea monosperma

(Lamk.) Taub. showed presence of protein and

triterpenes while absent in the powdered bark of

Butea monosperma(Lamk.) Taub. The powdered bark

and flowers of Butea monosperma (Lamk.) Taub.

showed presence of alkaloids and absent in leaf

powder of Butea monosperma (Lamk.) Taub.

Quantitative analysis

Quantitative analysis was carried for flavonoids,

alkaloids, phenols and saponins content inpowdered

plant parts (leaf, bark and flowers) of Butea

monosperma(Lamk.) Taub.The flavonoids content in

flowers samples collected was 130mg/gwas higher as

compared to that of leaf 70mg/g and bark was

50mg/g. Similarly the alkaloid content in flowers was

35mg/g and in the bark was 23mg/g. The saponin

content in different plant parts showed very less

difference. The saponin content in the flowers samples

was 66mg/g, in leaf 29mg/g, and bark 38mg/g. The

phenol content in bark 1.26mg/g of catechol, samples

was higher than the flowers 0.91mg/g of catechol, and

leaf 0.83mg/g of catechol in samples (Table 1.2).

Method/Test Plant parts collected from Murbad Location

L B F

Flavonoids (mg/g)

70.00 ±0.02

50.00 ±0.01

130.00 ±0.02

Alkaloids (mg/g)

0.00 23.00

±0.001 35.00

±0.002 Phenols (mg/g)

0.83 ±0.25

1.26 ±0.37

0.91 ±0.21

Saponins (mg/g)

29.00 ±0.001

38.00 ±0.002

66.00 ±0.002



a b c a b c a b c

254nm 366nm White light

Fig. 2.1: HPTLC fingerprint of flavonoids in Butea monosperma (Lamk.) Taub. plant parts (leaf, bark and flowers)

a b c a b c a b c

254nm 366nm White light

Fig. 2.2: HPTLC fingerprint of saponins in Butea monosperma (Lamk.) Taub. plant parts (leaf, bark and flowers).

HPTLC analysis

HPTLC is a valuable tool for the investigation of herbal

products with respect to different aspects of their

quality (Yamunadeviet al., 2012). HPTLC analysis was

tested to obtain reproductive peaks for flavonoids and

saponins. Flavonoids constitute one of the most

characteristic classes of compounds in plants. They

play important role in giving resistance to the plant

species (Mishra et al., 2012). The separation of

flavonoids was achieved using ethyl acetate-formic

acid-glacial acetic acid-water (100:11:11:26) as mobile

phase. Butea monosperma (Lamk.) Taub. Leafshowed 9

bands. Butea monosperma (Lamk.) Taub. bark showed

4 bands.Butea monosperma (Lamk.) Taub. flowers

showed 11 bandsYellow coloured fluorescent zone at

UV 366nm for flavonoids was observed in the

chromatogram and confirmed the presence of

flavonoids (Figure 2.1).

Saponins are known as non-volatile, surface active

compounds that are widely distributed in nature. They

Gupta et al., 2016


have a diverse range of medicinal properties

(Yamunadeviet al., 2012). The methanolic extracts of

leaf, bark and flowers of Butea monosperma (Lamk.)

Taub. showed presence of different types saponins

with different Rf values. The separation of saponins

was achieved using chloroform-acetic acid-methanol-

water (64:32:12:8) as mobile phase. Butea

monosperma (Lamk.) Taub. leaf showed 12 bands.

Butea monosperma (Lamk.) Taub.bark showed 9

bands. Butea monosperma (Lamk.) Taub. flowers

showed 11 bands. Blue-Violet and Yellow-Brown

coloured zones were observed from the

chromatogram and showed presence of saponins in

the sample (Figure 2.2).

CONCLUSION

Indian system such as Ayurveda and Siddha uses

majority of crude drugs that are of plant origin. It is

necessary to check the identity of the plant and

ascertain its quality before use. A detailed

pharmacognostic evaluation therefore is highly

essential prerequisite (Dhale, 2011). According to

World Health Organisation (WHO) the macroscopic

and microscopic description of a medicinal plant is the

first step towards establishing its identity and purity

and should be carried out before any tests are

undertaken (Anonymous, 2002).Thus the present

study,the stem of Butea monosperma (Lamk.) Taub.

Showsintrastelar and extrastelar secondary growth

with prominent diffuse porous wood and periderm

formation respectively. Deposition of yellow gummy

mass in pith, phloem and periderm is very

pronounced. Horse-shoe shaped or wedge shaped

stele which is a unique feature found in petiole of

Butea monosperma (Lamk.) Taub. These characters

may be considered as anatomical features for

identification of the plant.

It has been observed that the plant parts (leaf, bark

and flowers) of Butea monosperma (Lamk.) Taub.

showed presence of flavonoids, saponins and phenols

in preliminary and quantitative study. In quantitative

study the amount of flavonoids content in flowers of

Butea monosperma (Lamk.) Taub. was higher as

compared to that of leaf and bark. The developed

HPTLC fingerprint confirmed the presence of

flavonoids and saponins in plant parts of Butea

monosperma (Lamk.) Taub. The mobile phase was

found to be suitable for the study of flavonoids and

saponins separation. The developed phytochemical

analysis will help the manufacturers to distinguish the

adulterant and standardisation of herbal formulation.

In the present work, an attempt has been made to give

an insight on the anatomical features and

phytochemical analysisof various parts of the plant

Butea monosperma (Lamk.) Taub. This will help in

identifying the species and can be further used to

determine the botanical identity of herbal medicine.

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